• Title/Summary/Keyword: rtTA

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RTA Post-treatment of Thermal T${a_2}{O_5}$ Thin Films (열산화 T${a_2}{O_5}$박막에 미치는 RTA후처리의 영향)

  • Mun, Hwan-Seong;Lee, Jae-Seok;Han, Seong-Uk;Park, Sang-Gyun;Yang, Seung-Ji;Lee, Jae-Cheon;Park, Jong-Wan
    • Korean Journal of Materials Research
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    • v.3 no.3
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    • pp.310-315
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    • 1993
  • The effects of RT A treatment on the breakdown strengths were studied for tantalum pentoxide(${Ta_2}{O_5}$) films prepared by thermal oxidation of dc-sputtered Ta(400$\AA$) on p-type (100) Si wafer. While the relative dielectric constants of the RT A -treated specimens were not remarkably affected, the breakdown strengths of the RTA-treated specimens were greatly changed by RTA temperature and time. After the RTA treatment, the breakdown strengths of the specimens RTA-treated at the temperature below the crystallization temperature were increased to 5.4MV /cm, while those of the specimens RTA -treated at the temperature above it were decreased to 0.5MV /cm. RTA time-independence of the flat-bant voltage shift refleted that the RT A post-annealing effects on the breakdown strengths were not due to the interface reaction between the ${Ta_2}{O_5}$ layer and the Si substrate but, through the RBS analysis, to densification of the ${Ta_2}{O_5}$ films.

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Regulation of GFP Expression Using the Tetracycline Inducible Retroviral Vector System (Tetracycline Inducible Retrovirus Vector System에 의한 GFP 유전자의 발현 조절)

  • Koo Bon Chul;Kwon Mo Sun;Kim Teoan
    • Reproductive and Developmental Biology
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    • v.29 no.1
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    • pp.57-62
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    • 2005
  • One of the critical problems to be solved in transgenic animal production is uncontrollable constitutive expression of foreign genes, which usually results in serious physiological disturbances in the transgenic animal. To circumvent this problem, we constructed and tested two retrovirus vectors designed to express the GFP(green fluorescent protein) gene under the control of the tetracycline-inducible promoters. To maximize the GFP gene expression at turn-on state, WPRE(woodchuck hepatitis virus posttranscriptional regulatory element) sequence was introduced into the retrovirus vectors at downstream region of either the GFP gene or the sequence encoding rtTA(reverse tetracycline-controlled transactivator). Transformed cells were cultured in the medium supplemented with or without doxycycline(tetracycline derivative) for 48 hours, and induction efficiency was measured by comparing the GFP gene expression level using fluorometry and western blotting. Higher GFP expression was observed from the vector carrying the WPRE sequence at 3' side of the GFP gene, while tighter expression control(up to 20 fold) was obtained from the vector in which the WPRE sequence was placed at 3' side of rtTA sequence. The resulting tetracycline inducible vector system may be used in transgenic animal production and gene therapy.

Efficient Control of Human G-CSF Gene Expression in the Primary Culture Cell using a FIV-Tet-On Vector System (FIV-Tet-On Vector System을 이용한 hG-CSF 유전자의 효율적인 발현 조절)

  • Kwon, Mo-Sun;Koo, Bon-Chul;Kim, Te-Oan
    • Reproductive and Developmental Biology
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    • v.31 no.3
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    • pp.153-159
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    • 2007
  • In this study, using FIV-based lentivirus vector system, we tried to express hG-CSF in tetracycline-controllable manner. hG-CSF influences the proliferation, differentiation, and survival of cells in the neutrophil lineage. To enhance stability and translation of hG-CSF transcript, WPRE sequence was also introduced into FIV-Tet-On vector at downstream region of either the hG-CSF gene or the sequence encoding rtTA. Primary culture cells (CEF, chicken embryonic fibroblast; PFF, procine fetal fibroblast) infected with the recombinant FIV were cultured in the medium supplemented with or without doxycycline for 48 hours, and induction efficiency was measured by comparing the hG-CSF gene expression level using quantitative real-time PCR, Western blot and ELISA. Higher hG-CSF expression and tighter expression control were observed from the vector in which the WPRE sequence was placed at downstream of the hG-CSF (in CEF) or rtTA (in PEE) gene. This FIV-Tet-On vector system may be helpful in solving serious physiological disturbance problems which has continuously hampered successful production of transgenic animals and gene therapy.

Regulation of hPTH Expression In Virto Using the Tetracycline Inducible Retrovirus Vector System (Tetracycline Inducible Retrovirus Vector System을 이용한 In Vitro에서의 인간 부갑상선 호르몬의 발현 조절)

  • Koo, Bon-Chul;Kwon, Mo-Sun;Kim, Te-Oan
    • Reproductive and Developmental Biology
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    • v.30 no.3
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    • pp.157-162
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    • 2006
  • Endogenous 84 amino acid parathyroid hormone(PTH) is synthesized as a pre-pro hormone by the chief cells of the parathyroid glands. Physiological actions of PTH include regulation of bone metabolism, renal tubular reabsorption of calcium and phosphate, and intestinal calcium absorption. In addition, PTH stimulates new bone formation by extraordinary stimulation of osteoblastic activity and decreasing calcium excretion by the kidney. In this study, we constructed and tested retrovirus vectors designed to express the human parathyroid hormone(hPTH) gene under the control of the tetracycline-inducible promoters. To increase the hPTH gene expression at turn-on state, woodchuck hepatitis virus posttranscriptional regulatory element(WPRE) sequence was also introduced into retrovirus vector at downstream region of either the hPTH gene or the sequence encoding reverse tetracycline-controlled transactivator(rtTA). Transformed primary culture cells(porcine fetal fibroblast, PFF, chicken embryonic fibroblast, CEF) were cultured in the medium supplemented with or without doxycycline(tetracycline derivative) for 48 hours, and induction efficiency was measured by comparing the hPTH gene expression level using two step RT-PCR and ELISA Higher hPTH expression($3{\tims}10^4\;pg/ml,\;5.3{\times}10^4\;pg/ml$) and tighter expression control(up to 8 fold) were observed from the vector in which the WPRE sequence was placed at downstream of the hPTH gene. The resulting tetracycline inducible vector system may be helpful in solving serious physiological disturbance problems which have been a major obstacle in successful production of transgenic animals.

Stabilization of $Pb(Mg_{1/3}Ta_{2/3})O_3$ thin film by a thin $PbTiO_3$ seed layer and characterization of electric properties ($PbTiO_3$ 씨앗층을 이용한 $Pb(Mg_{1/3}Ta_{2/3})O_3$ 박막의 상안정화와 전기적 특성평가)

  • 김태언;유창준;문종하;김진혁
    • Proceedings of the Materials Research Society of Korea Conference
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    • 2003.03a
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    • pp.211-211
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    • 2003
  • PbTiO$_3$ 씨앗층을 이용하여 완화형 강유전체 Pb(Mg$_{1}$3/Ta$_{2}$3/)O$_3$ (PMT) 박막의 페로브스카이트 상안정화와 열처리 조건에 따른 미세구조변화, 이에 따른 전기적 특성 변화에 관하여 조사하였다. PbTiO$_3$ 박막을 스핀코팅법으로 3000 rpm에서 20초간(111) 방향으로 배향된 Pt / Ti / SiO$_2$/ Si 기판에 증착하여 안정화된 페로브스카이트 박막을 얻었다. 이렇게 제조된 PbTiO$_3$를 Buffer 층으로 사용하고 그 위에 Pb(Mg$_{1}$3/Ta$_{2}$3/)O$_3$를 박막을 Spin coating방법으로 증착한 후, 급속열처리 방법(RTA)으로 550- $650^{\circ}C$ 사이에서 열처리하였다. 제조된 박막의 열처리 온도에 따른 미세구조 변화와 결정성을 XRD, SEM, TEM으로 분석하였고 박막의 저온 강유전 특성을 RT66A를 이용하여 평가하였다. Pb(Mg$_{1}$3/Ta$_{2}$3/)O$_3$ 박막의 경우 씨앗층이 없는 경우에는 pyrochlore상이 주상이었지만 씨앗층을 사용한 경우 페로브스카이트 상이 주상임을 확인하였고 열처리 온도가 증가할수록 페로브스카이트상의 상대적 양이 증가함을 확인하였다. 미세구조와 상의 변화에 따른 전기적 특성 변화에 관하여 자세하게 논의할 것이다.

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Induction of cancer cell-specific death via MMP2 promoterdependent Bax expression

  • Seo, Eun-Jeong;Kim, Se-Woon;Jho, Eek-hoon
    • BMB Reports
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    • v.42 no.4
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    • pp.217-222
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    • 2009
  • Controlled gene expression in specific cells is a valuable tool for gene therapy. We attempted to determine whether the lentivirus-mediated Tet-On inducible system could be applied to cancer gene therapy. In order to select the genes that induce cancer cell death, we compared the ability of the known pro-apoptotreic genes, Bax and tBid, and a cell cycle inhibitor, p21cip1/waf1, and determined that Bax was the most effective. For the cancer cell-specific expression of $rtTA2^S$-M2, we tested the matrix metalloproteinase-2 (MMP-2) promoter and determined that it is highly expressed in cancer cell lines, including SNU475 cells. The co-transduction of two lentiviruses that contain sequences for TRE-Bax and $rtTA2^S$-M2, the expression of which is controlled by the MMP-2 promoter, resulted in the specific cell death of SNU475, whereas other cells with low MMP-2 expression did not evidence significant cell death. Our data indicate that the lentivirus-mediated Tet-On system using the cancer-specific promoter is applicable for cancer gene therapy.

Cell Type-Specific and Inducible PTEN Gene Silencing by a Tetracycline Transcriptional Activator-Regulated Short Hairpin RNA

  • Wang, Shan;Wang, Ting;Wang, Tao;Jia, Lintao
    • Molecules and Cells
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    • v.38 no.11
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    • pp.959-965
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    • 2015
  • Inducible and reversible gene silencing in desired types of cells is instrumental for deciphering gene functions using cultured cells or in vivo models. However, efficient conditional gene knockdown systems remain to be established. Here, we report the generation of an inducible expression system for short hairpin RNA (shRNA) targeted to PTEN, a well-documented dual-specificity phosphatase involved in tumor suppression and ontogenesis. Upon induction by doxycycline (DOX), the reverse tetracycline transcriptional activator (rtTA) switched on the concomitant expression of GFP and a miR-30 precursor, the subsequent processing of which released the embedded PTEN-targeted shRNA. The efficacy and reversibility of PTEN knockdown by this construct was validated in normal and neoplastic cells, in which PTEN deficiency resulted in accelerated cell proliferation, suppressed apoptosis, and increased invasiveness. Transgenic mice harboring the conditional shRNA-expression cassette were obtained; GFP expression and concurrent PTEN silencing were observed upon ectopic expression of rtTA and induction with Dox. Therefore, this study provides novel tools for the precise dissection of PTEN functions and the generation of PTEN loss of function models in specific subsets of cells during carcinogenesis and ontogenesis.

Electrical and Chemical Properties of ultra thin RT-MOCVD Deposited Ti-doped $Ta_2O_5$

  • Lee, S. J.;H. F. Luan;A. Mao;T. S. Jeon;Lee, C. h.;Y. Senzaki;D. Roberts;D. L. Kwong
    • JSTS:Journal of Semiconductor Technology and Science
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    • v.1 no.4
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    • pp.202-208
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    • 2001
  • In Recent results suggested that doping $Ta_2O_5$ with a small amount of $TiO_2$ using standard ceramic processing techniques can increase the dielectric constant of $Ta_2O_5$ significantly. In this paper, this concept is studied using RTCVD (Rapid Thermal Chemical Vapor Deposition). Ti-doped $Ta_2O_5$ films are deposited using $TaC_{12}H_{30}O_5N$, $C_8H_{24}N_4Ti$, and $O_2$ on both Si and $NH_3$-nitrided Si substrates. An $NH_3$-based interface layer at the Si surface is used to prevent interfacial oxidation during the CVD process and post deposition annealing is performed in $H_2/O_2$ ambient to improve film quality and reduce leakage current. A sputtered TiN layer is used as a diffusion barrier between the Al gate electrode and the $TaTi_xO_y$ dielectric. XPS analyses confirm the formation of a ($Ta_2O_5)_{1-x}(TiO_2)_x$ composite oxide. A high quality $TaTi_xO_y$ gate stack with EOT (Equivalent Oxide Thickness) of $7{\AA}$ and leakage current $Jg=O.5A/textrm{cm}^2$ @ Vg=-1.0V has been achieved. We have also succeeded in forming a $TaTi_x/O_y$ composite oxide by rapid thermal oxidation of the as-deposited CVD TaTi films. The electrical properties and Jg-EOT characteristics of these composite oxides are remarkably similar to that of RTCVD $Ta_2O_5, suggesting that the dielectric constant of $Ta_2O_5$ is not affected by the addition of $TiO_2$.

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Properties of SBT Thin Film Synthesized by Self-seed Layer Method (Self-seed layer를 이용하여 증착한 SBT박막의 특성)

  • Kim, Hyung-Sub;Hwang, Dong-Hyun;Yoon, Ji-Un;Son, Young-Gook
    • Journal of the Korean Vacuum Society
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    • v.16 no.3
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    • pp.215-220
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    • 2007
  • Thin films of $SBT(SrBi_2Ta_2O_9)$ having $Pt/SBT/Seed/Pt/Ti/SiO_2/Si$ structure were fabricated using self-seed layer method by R.F. Magnetron sputter. Self-seed layers were deposited at room temperature and $600^{\circ}C$, which had 30 nm thickness. To investigate crystallization of self-seed layer we characterized by XRD after various heat treatment. And we characterized the crystallinity and electrical properties of SBT on self-seed layer after various heat treatment.

Biocompatibility of Ti-8Ta-3Nb alloy with fetal rat calvarial cells (백서 태자 두개관세포에서 Ti-8Ta-3Nb 합금의 생체적합성)

  • Cho, In-Goo;Cui, De-Zhe;Kim, Young-Joon;Lee, Kyung-Ku;Lee, Doh-Jae
    • Journal of Periodontal and Implant Science
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    • v.36 no.4
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    • pp.849-861
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    • 2006
  • 타이타늄은 기계적 특성이 우수하고 생체적합성이 뛰어나 의료용 장비의 주 재료로 사용되고 있으며 타이타늄 보다 기계적 특성이 더 우수한 타이타늄 합금들(주로 Ti-6Al-4V와 Ti-6Al-7Nb 합금)도 개발되어 치과와 의료용 임플란트로 사용되고 있다. 그러나 타이타늄 합금 성분들 중 알루미늄 (aluminum)과 바나디움(vanadium)은 인체에 노출되면 세포손상과 신경계에 문제를 일으킬 수 있다. 따라서 인체에 독성이 없으면서 기계적 성질과 생체적합성이 우수한 타이타늄 합금의 개발이 필요하다. 최근 인체에 독성이 없는 성분들이 함유된 새로운 ${\beta}$ - 형태의 타이타늄 합금들이 개발되고 있는데, ${\beta}$ - 타이타늄 합금은 그 기계적 성질이 기존의 ${\alpha}+{\beta}$ 타이타늄 합금에 비해 우수하다고 알려져 있다. 최근 새로운 ${\beta}$ - 타이타늄 합금이 전남대학교 부설 타이타늄 연구소에서 개발되었다. 이 연구는 새로 개발된 ${\beta}$ - 타이타늄 합금의 생채 적합성을 세포 증식도, 알카리 인산 분해 효소 활성과 유전자 증폭을 통해 알아보고자 하였다. 그 결과는 다음과 같다; 1. Titanium-6aluminum-4vanadium (Ti-6Al-4V) 합금 표면애서의 세포 증식율은 Titanium-Titanium8Tantalum-3Niobium (Ti-8Ta-3Nb) 합금과 순수 타이타늄 표면에 비해 유의하게 낮았다(p<0.00l). Ti-8Ta-3Nb 합금 표면에서의 증식도는 순수 타이타늄 표면과 유사하였다. 2. Ti-8Ta-3Nb 합금과 순수 타이타늄에서 배양된 세포이 알카리 인산 분해 효소의 활성도는 Ti-6Al-4V 합금에서의 것보다 유의하게 높았다 (p<0.001). 3. 유전자 증폭 분석 결과, Ti-8Ta-3Nb 합금과 순수 타이타늄에서 collagen type I과 bone sialoprotein mRNA 가 유사한 수준으로 발현되었다. 이상의 결과는 생체 적합성 측면에서 Ti-8Ta-3Nb 합금과 순수 타이타늄의 차이가 없음을 보여주며 따라서 Ti-8Ta-3Nb 합금이 의학 및 치의학 영역에서 새로운 임프란트 재료로 사용될 수 있음을 의미한다.